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目的:探讨上皮膜蛋白1(epithelial membrane protejn-1,EMP1)在人结直肠癌(colorectal carcinoma,CRC)组织中的表达水平及其过表达对CRC SW-480细胞增殖、凋亡和侵袭的影响。方法:免疫组织化学、Western blotting方法检测63例CRC组织及31例癌旁组织中EMP1的表达水平,分析EMP1表达与CRC临床病理参数的关系。慢病毒介导将pLenti6-EMP1质粒转染SW-480细胞,建立EMP1过表达细胞,转染plenti6/V5-DEST空白质粒为对照。Real-time PCR及Western blotting检测转染后SW-480细胞株中EMP1的表达,MTT、流式细胞术及Transwell实验分别检测EMP1过表达对SW-480细胞增殖、凋亡及侵袭能力的影响。结果:EMP1蛋白在人CRC组织的表达显著低于癌旁组织(0.257±0.022 vs 0.863±0.086,P<0.05),并且其表达水平在不同T分期、有无淋巴结转移、临床分期以及组织分级组间表达有显著差异(P<0.05),而与患者年龄、性别、肿瘤部位无关(P>0.05)。成功构建EMP1过表达的LeEMP1细胞。与LeEmpty细胞相比,LeEMP1细胞的增殖能力显著下降[(60.94±4.04)%vs(100.00±0.00)%,P<0.05],凋亡率显著升高[(12.10±1.30)%vs(3.10±0.60)%,P<0.05]、侵袭转移能力显著降低[穿膜细胞数:(87.00±12.00)vs(178.00±21.00)个,P<0.05]。LeEMP1细胞Caspase-9表达显著高于LeEmpty细胞(0.764±0.073 vs 0.231±0.029,P<0.05),VEGFC表达显著降低(0.185±0.022 vs 0.663±0.065,P<0.05)。结论:人CRC组织中EMP1蛋白表达明显减低,过表达EMP1能够抑制CRC SW-480细胞的恶性生物学行为,其可能通过调控Caspase-9和VEGFC的表达来发挥作用。
Objective: To investigate the expression of epithelial membrane protein 1 (EMP1) in human colorectal carcinoma (CRC) and its effect on the proliferation, apoptosis and invasion of CRC SW-480 cells . Methods: Immunohistochemistry and Western blotting were used to detect the expression of EMP1 in 63 CRC tissues and 31 adjacent normal tissues. The relationship between EMP1 expression and CRC clinicopathological parameters was analyzed. Lentivirus-mediated transfection of pLenti6-EMP1 plasmid into SW-480 cells, establishment of EMP1 overexpression cells, transfected plenti6 / V5-DEST blank plasmid as a control. The expression of EMP1 in SW-480 cells was detected by Real-time PCR and Western blotting. The effects of EMP1 overexpression on the proliferation, apoptosis and invasion of SW-480 cells were detected by MTT, flow cytometry and Transwell assay. Results: The expression of EMP1 protein in human CRC tissues was significantly lower than that in adjacent non-cancerous tissues (0.257 ± 0.022 vs 0.863 ± 0.086, P <0.05), and the expression level of EMP1 protein in different T stages, with or without lymph node metastasis, clinical stage and histological grading There was significant difference between the two groups (P <0.05), but not with age, gender and tumor location (P> 0.05). EMP1 overexpressed LeEMP1 cells were successfully constructed. Compared with LeEmpty cells, the proliferation of LeEMP1 cells was significantly decreased [(60.94 ± 4.04)% vs (100.00 ± 0.00)%, P <0.05], and the apoptosis rate was significantly increased [(12.10 ± 1.30)% vs 0.60)%, P <0.05]. The invasion and metastasis abilities decreased significantly (the number of transmembrane cells: (87.00 ± 12.00) vs (178.00 ± 21.00), P <0.05]. The expression of Caspase-9 in LeEMP1 cells was significantly higher than that in LeEmpty cells (0.764 ± 0.073 vs 0.231 ± 0.029, P <0.05), and the expression of VEGFC was significantly decreased (0.185 ± 0.022 vs 0.663 ± 0.065, P <0.05). CONCLUSION: The expression of EMP1 protein in human CRC tissue is significantly reduced. Overexpression of EMP1 can inhibit the malignant biological behavior of CRC SW-480 cells, which may play a role in regulating the expression of Caspase-9 and VEGFC.